Karyotypes of Pneumocystis carinii from Korean rats.

Molecular karyotyping was applied to Pneumocystis carinii(Pc) from two strains of experimental rats, Sprague Dawley(SD) and Fisher(F), in Korea. Field inversion gel electrophoresis and contour clamped homogeneous electric field electrophoresis resolved 15 chromosomal bands from the Pc. The size of the bands was estimated 270kb to 684kb from SD rats, and 273kb to 713 kb from F rats. The bands of 283 kb from SD rats and of 273 kb from F rats stained more brightly suggesting duplicated bands. Total number of chromosomes was at least 16, and total genomic size was estimated 7 x 10(6) bp. All of the bands from F rats hybridized to the probe of repeated DNA sequences of Pc and the band of 448 kb size was proved to contain rDNA sequences, but Pc. chromosome bands from SD rats showed no reactions to the probes. The 2 different karyotypes of P. carinii from 2 strains of rats were maintained consistently for 2 years.     

Purification of monomeric agmatine iminohydrolase from soybean

Agmatine iminohydrolase (EC 3.5.3.12) was purified to homogeneity from the cytosol of soybean (Glycine max) axes by chromatographic separations on Sephadex G-25, Bio-rex 70, and agmatine-affinity columns. The enzyme was homogeneous by the criteria of analytical gel electrophoresis. Molecular weights estimated by Sephadex G-100 gel and sodium dodecyl sulfate polyacrylamide gel electrophoresis were 70,000, indicating that the soybean axes enzyme is a monomer, in contrast to the dimeric enzymes from corn and rice. The isoelectric point determined by gel electrofocusing was 7.5, higher than that of the corn enzyme (4.7). The optimal pH and temperature for activity were 6.5 and 50 degrees C, respectively. The enzyme has high specificity for agmatine, and the Km for agmatine was 2.5 x 10(-3) molar. The enzyme was sensitive to Cu2+ and also was inhibited by p-hydroxymercuribenzoate.     

A linkage map of distal mouse chromosome 12

To refine the linkage map of distal mouse Chromosome 12, we have identified DNA restriction fragment variants associated with a creatine kinase gene (Ck-3), the Akt proto-oncogene, an Abelson proviral integration site (D12N1), and the immunoglobulin heavy chain V_H3609 variable region family (Igh-V36). The patterns of inheritance of these markers in backcross progeny and recombinant inbred mouse strains allowed their localization with respect to previously mapped genes to yield the linkage map: Aat-15.8 cM-Ck-3-0.9 cM-(Crip, Akt, Igh-C)-0.3 cM-(D12N1, Igh-V). This map confirms genetically the localization of the Igh-V gene complex distal to Igh-C on the chromosome. It differs from previous maps in placing D12N1 distal to Igh-C, and in suggesting that the Igh-V gene complex spans less than one centiMorgan (cM).Other DNA sequence variants detected with the creatine kinase probe allowed definition of four additional genetic loci: Ck-1 near Lmyc-1 on Chromosome 4; Ck-2 between Upg-1 and Hprt-ps1 (D17Rp10) on distal Chromosome 17; Ck-4 near Mpmv-17 and Mls-3 on Chromosome 16; and Ck-5 near Hba on Chromosome 11.     

Coupling of proadipocyte growth arrest and differentiation. II. A cell cycle model for the physiological control of cell proliferation

Experimental evidence is presented that supports a cell cycle model showing that there are five distinct biological processes involved in proadipocyte differentiation. These include: (a) growth arrest at a distinct state in the G1 phase of the cell cycle; (b) nonterminal differentiation; (c) terminal differentiation; (d) loss of the differentiated phenotype; and (e) reinitiation of cell proliferation. Each of these events is shown to be regulated by specific human plasma components or other physiological factors. At two states designated GD and GD', coupling of growth arrest and differentiation is shown to occur. We propose that these mechanisms for the coupling of growth arrest and differentiation are physiologically significant and mimic the regulatory processes that control stem cell proliferation in vivo.     

Immobilization of enzymes on activated carbon: selection and preparation of the carbon support.

Based upon its superior catalytic activity for H2O2 decomposition, a bituminous coal-based activated carbon was selected for investigations of pretreatment and enzyme immobilization methods. Pretreatments considered include acid washing, exposure to strong oxidizing agents, contact with concentrated peroxide solutions, nitration and amination, isothiocyanate derivatization, silanization, and stearic acid coating. Effects of these pretreatments on morphology and trace-metal content of the carbon pellets have been studied using scanning electron microscopy and dispersive analysis of x rays. Immobilization of glucoamylase by adsorption, glutaraldehyde crosslinking, and covalent attachment to carbon activated by water-soluble diimide or diazotization have been examined. These different enzyme-carbon catalysts have been characterized by their enzyme loading, enzyme activity, catalytic activity for H2O2 decomposition, or combinations of these measures of performance.     

Spectrophotometric determination of bromopyruvate by reaction with 2-nitro-5-thiobenzoic acid

A spectrophotometric method for determination of bromopyruvate, based on the reaction with 2-nitro-5-thiobenzoic acid, is described. The reaction is complete in 30 min at room temperature in 0.1 m Tris-MES, 1 mm EDTA, pH 8.0. The method is sensitive to at least 1 × 10^?5m bromopyruvate. Reagents are stable, easy to prepare, and specific for β-halopyruvate. Bromopyruvate solutions must be prepared fresh daily. Solutions of bromopyruvate at pH 8.0 and 23°C have a half-life of 3 hr.     

Role of PCK2 in the proliferation of vascular smooth muscle cells in neointimal hyperplasia

Vascular smooth muscle cell (VSMC) proliferation is a hallmark of neointimal hyperplasia (NIH) in atherosclerosis and restenosis post-balloon angioplasty and stent insertion. Although numerous cytotoxic and cytostatic therapeutics have been developed to reduce NIH, it is improbable that a multifactorial disease can be successfully treated by focusing on a preconceived hypothesis. We, therefore, aimed to identify key molecules involved in NIH via a hypothesis-free approach. We analyzed four datasets ({"type":"entrez-geo","attrs":{"text":"GSE28829","term_id":"28829"}}GSE28829, {"type":"entrez-geo","attrs":{"text":"GSE43292","term_id":"43292"}}GSE43292, {"type":"entrez-geo","attrs":{"text":"GSE100927","term_id":"100927"}}GSE100927, and {"type":"entrez-geo","attrs":{"text":"GSE120521","term_id":"120521"}}GSE120521), evaluated differentially expressed genes (DEGs) in wire-injured femoral arteries of mice, and determined their association with VSMC proliferation in vitro. Moreover, we performed RNA sequencing on platelet-derived growth factor (PDGF)-stimulated human VSMCs (hVSMCs) post-phosphoenolpyruvate carboxykinase 2 (PCK2) knockdown and investigated pathways associated with PCK2. Finally, we assessed NIH formation in Pck2 knockout (KO) mice by wire injury and identified PCK2 expression in human femoral artery atheroma. Among six DEGs, only PCK2 and RGS1 showed identical expression patterns between wire-injured femoral arteries of mice and gene expression datasets. PDGF-induced VSMC proliferation was attenuated when hVSMCs were transfected with PCK2 siRNA. RNA sequencing of PCK2 siRNA-treated hVSMCs revealed the involvement of the Akt-FoxO-PCK2 pathway in VSMC proliferation via Akt2, Akt3, FoxO1, and FoxO3. Additionally, NIH was attenuated in the wire-injured femoral artery of Pck2-KO mice and PCK2 was expressed in human femoral atheroma. PCK2 regulates VSMC proliferation in response to vascular injury via the Akt-FoxO-PCK2 pathway. Targeting PCK2, a downstream signaling mediator of VSMC proliferation, may be a novel therapeutic approach to modulate VSMC proliferation in atherosclerosis.     

北京山区黄栌叶片水分和光能利用效率季节变化及影响因子

未来气候变化将影响光合环境资源供给,尤其是水分和光能。为深入了解植物对气候变化的适应性,使用LI-6800便携式光合仪,于2021年5—10月份(完全展叶期)测定了北京山区广布灌木黄栌(Cotinus coggygria)叶片的光响应曲线,分析其水分利用效率(WUE=最大净光合速率[P_nmax]/气孔导度[g_s])和光能利用效率(LUE)的季节变化特征及影响因子。结果显示：黄栌叶片WUE在5—6月份呈下降趋势,7—10月份比较稳定;LUE在5—7月份呈上升趋势,8—10月份比较稳定。WUE和LUE的生长季平均值分别为98.25μmol/mol和0.06 mol/mol,变异系数分别为22%和17%,两者呈负相关(R²=0.86;P<0.01)。环境因子中,WUE和LUE主要受土壤含水量(SWC)影响,WUE随SWC增加呈线性降低趋势,而LUE随SWC增加呈线性增加趋势。SWC每增加0.1 m³/m³,P_nmax和g_s分别线性增加...     

六种海桑属红树植物根际真菌特征及其影响因子

真菌多样性是植物根际生态系统的重要构成与植物健康稳定的重要指标。海桑属是红树林的先锋物种,采用真菌ITS1区高通量测序方法,分析了六种海桑属红树根际真菌的组成和多样性,结合土壤理化性质探讨影响不同植物根际真菌群落组成差异的因素。结果显示,根际真菌隶属于7门、96科、155属,子囊菌门作为优势菌门在海桑属不同红树中相对丰度无显著差异,都超过27%,但次优势的担子菌门丰度含量有差异;属水平上,优势菌属的丰度含量不同,曲霉属在卵叶海桑的丰度最高(29.57%),在海南海桑最低(3.47%)。六种红树植物根际存在特有的代表类群,如无瓣海桑的马拉色菌(9.31%)和毛腐菌属(10.05%),海南海桑中的Talaromyces(19.61%)和Acremonium(13.58%)。比较多样性指数Simpson和Shannon,发现拟海桑是六种植物中丰度最高的,卵叶海桑最低。RDA分析发现子囊菌门与全磷含量呈显著负相关,担子菌门与速效钾呈明显正相关。六种海桑属红树植物根际核心物种分析表明,优势真菌类群曲霉属和一些低丰度的真菌类群,通过降解有机质参与碳循环,对根际土壤生态系统的稳定起重要作用。六种海桑...